We propose to identify and characterize proteins in innermost layer of Ixodes scapularis and Amblyomma americanum tick cement cones. Currently there are 14 reportable human tick borne disease (TBD) agents are recognized in the United States with 13 of these transmitted via hard tick bites (http://www.cdc.gov/ticks/diseases/). Hard tick species are adapted to feed for many days. Majority of hard ticks secrete an adhesive substance known as tick cement that allows ticks to firmly anchor onto host skin, and prevent them from being groomed off their feeding site by their host. The tick cement cone has two layers: the innermost layer, which is deposited within 5-30min of the tick starting the feeding process, and the outermost layer, which begins to be deposited one day after the tick has started the feeding process. Deposition of the innermost layer precedes major tick feeding events including transmission of major human tick borne diseases such as the Lyme disease agent. Thus, our assumptions are that preventing functions of proteins that form the innermost layer of the tick cement cone will stop tick feeding before transmission of major human disease agents starts. There are two specific aims in this application. First is to identify proteins in I. scapularis and A. americanum tick cement cones, and second is to verify significance in tick attachment and/or completion of tick feeding. Of the 14 reportable human TBD agents, in the United States, a combined 9 are vectored by I. scapularis and A. americanum (http://www.cdc.gov/ticks/diseases/). Thus proposed discovery of I. scapularis and A. americanum tick cement proteins will have broader impacts toward development of novel methods to improve public health in the USA.